TECHNIQUES AND METHODS

Selection and Validation of Reference Genes for Quantitative RT-PCR Analysis in Neolamarckia cadamba

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  • Guangdong Key Laboratory for Innovative Development and Utilization of Forest Plant Germplasm, College of Forestry and Landscape Architecture, South China Agricultural University, Guangzhou 510642, China

Received date: 2018-01-03

  Online published: 2018-04-26

Abstract

In this study, root, shoot, leaf, flower, fruit, peel and cambium tissue of Neolamarckia cadamba was sampled to analyze the expression of 43 candidate reference genes in 21 housekeeping gene families, such as ACT, CAC, CYP, and EF1α, by RT-qPCR. The software geNorm, NormFinder and BestKeeper were used to analyze expression stability of these candidate reference genes in the seven different tissues. geNorm analysis revealed that the stability of the UPL gene was the highest (M=0.443) and the stability of UBQ was the lowest (M=2.859). NormFinder analysis revealed that the stability of UPL was the highest (E=0.223), UBQ was the lowest (M=4.759). BestKeeper analysis revealed that the standard deviation of UPL (SD=0.513) was the lowest. These findings suggest that UPL has the highest stability, and UBQ has the lowest stability. UPL gene could be selected as an internal reference gene for analysis of gene expression among different tissues of N. cadamba.

Cite this article

Zhang Deng, Li Jingjian, Zhang Mengjie, Bao Yutao, Yang Xiao, Xu Wuyun, Ouyang Kunxi, Chen Xiaoyang . Selection and Validation of Reference Genes for Quantitative RT-PCR Analysis in Neolamarckia cadamba[J]. Chinese Bulletin of Botany, 2018 , 53(6) : 829 -839 . DOI: 10.11983/CBB18003

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